Induction of 72-kDa inducible heat shock protein (HSP72) in cultured rat astrocytes after energy depletion.
Protein synthesis is important in the readaptive processes for cultured astrocytes after hypoxia and subsequent reoxygenation. We have identified 72-kDa inducible heat shock protein (HSP72) as a major stress protein in reoxygenated astrocytes. To assess the mechanism for reoxygenation-mediated induction of HSP72, a reporter gene that consists of a human HSP promoter fused to the luciferase gene was transfected into cultured astrocytes. Analysis of cellular energy nucleotides showed an increase of the ADP/ATP ratio after reoxygenation, which synchronized with activation of the HSP promoter. Activation of the HSP promoter was also observed after an addition of iodoacetic acid to hypoxic astrocytes, which reached the maximum when the ADP/ATP ratio reached 50%, but further decline in the energy profile caused inactivation of this promoter. Inhibition of protein synthesis after reoxygenation resulted in temporary restoration of the energy profile and suppression of the DNA binding activity of the heat shock factor. Addition of quercetin greatly decreased the [3H]leucine incorporation in the polysome fraction without any effect on the mature mRNA formation. These data suggest that the energy depletion in reoxygenation triggers induction of HSP72 after reoxygenation, which may act as a pivotal mediator in the stress response of reoxygenated astrocytes by facilitating protein synthesis.
Imuta
N
N
Department of Neurology, Osaka University Medical School, Suita, Japan.
Ogawa
S
S
Maeda
Y
Y
Kuwabara
K
K
Hori
O
O
Ueda
H
H
Yanagihara
T
T
Tohyama
M
M
D000227:Adenine Nucleotides / Q000378:metabolism*; D000332:Aerobiosis; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004734:Energy Metabolism / Q000187:drug effects / Q000502:physiology*; D017930:Genes, Reporter; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000096:biosynthesis*; D006801:Humans; D008156:Luciferases / Q000096:biosynthesis; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011794:Quercetin / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D014452:Ubiquitins / Q000096:biosynthesis
D000227:Adenine Nucleotides; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; D014452:Ubiquitins; D011794:Quercetin; D008156:Luciferases